LOCUS       sequence-431433-        2952 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fragmid fragment: (N' terminus) ZIM3 KRAB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2952)
  AUTHORS   McGee AV, Liu YV, Griffith AL, Szegletes ZM, Wen B, Kraus C, Miller 
            NW, Steger RJ, Escude Velasco B, Bosch JA, Zirin JD, Viswanatha R, 
            Sontheimer EJ, Goodale A, Greene MA, Green TM, Doench JG
  TITLE     Modular vector assembly enables rapid assessment of emerging CRISPR 
            technologies.
  JOURNAL   Cell Genom. 2024 Mar 13;4(3):100519. doi: 
            10.1016/j.xgen.2024.100519.
  PUBMED    38484704
REFERENCE   2  (bases 1 to 2952)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell 
            Genom."; date: "2024-03-13"; volume: "4(3):100519. doi"; pages: " 
            10.1016/j.xgen.2024.100519"
COMMENT     Sequence Label: AA088
FEATURES             Location/Qualifiers
     source          1..2952
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     regulatory      444..453
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             489..509
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(862..878)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(862..878)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(875..897)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    886..902
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(910..916,917..934,935..940))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    955..976
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1093..1110)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1264..1852)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1344..1363)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2030..2839)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2753..2772
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2831..2850
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagagaatt cgagctcggt acctcgcgaa
      421 tacatctaga tgaagacgta ggagccacca tggccaaacg gacagccgac ggaagcgagt
      481 tcgagtcacc aaagaagaag cggaaagtca gtggcggcag cagcggcaac aacagccagg
      541 gcagagtgac cttcgaggac gtgaccgtga acttcaccca gggcgagtgg cagagactga
      601 accccgagca gagaaacctg tacagagatg tgatgctgga gaactacagc aacctggtga
      661 gcgtgggcca gggcgagacc accaagcccg acgtgatcct gagactggag cagggcaagg
      721 agccctggct ggaggaggag gaggtgctgg gcagcggcag agccgagtct ggaggatcta
      781 gcggaggatc cggcagcagt cttcatcgga tcccgggccc gtcgactgca gaggcctgca
      841 tgcaagcttg gtgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
      901 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
      961 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     1021 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     1081 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     1141 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     1201 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     1261 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     1321 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     1381 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     1441 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     1501 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     1561 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     1621 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     1681 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     1741 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     1801 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     1861 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     1921 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     1981 taaatcaagc ccaatctgaa taatgttaca accaattaac caattctgat tagaaaaact
     2041 catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt
     2101 gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa
     2161 gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc
     2221 cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg
     2281 agaatggcaa aagtttatgc atttctttcc agacttgttc aacaggccag ccattacgct
     2341 cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga
     2401 gacgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc
     2461 gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata
     2521 cctggaatgc tgtttttccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac
     2581 ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca
     2641 tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg
     2701 catcgggctt cccatacaag cgatagattg tcgcacctga ttgcccgaca ttatcgcgag
     2761 cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc ctcgacgttt
     2821 cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca gacagtttta
     2881 ttgttcatga tgatatattt ttatcttgtg caatgtaaca tcagagattt tgagacacgg
     2941 gccagagctg ca
//
